Structural basis for DNA unwinding at forked dsDNA by two coordinating Pif1 helicases

Structural basis for DNA unwinding at forked dsDNA by two coordinating Pif1 helicases
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两个协调 Pif1 解旋酶在分叉 dsDNA 上解旋 DNA 的结构基础

DOI:
10.1038/s41467-019-13414-9
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发表时间:
2019-11
影响因子:
16.6
通讯作者:
Haiwei Song
Haiwei Song
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Nannan Su;Alicia K. Byrd;Sakshibeedu R. Bharath;Olivia Yang;Yu Jia;Xuhua Tang;Taekjip Ha;Kevin D. Raney;Haiwei Song

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Pif 1在维持基因组稳定性和优先解旋叉状dsDNA中起着多种作用,但Pif 1解旋叉状dsDNA的机制仍然难以捉摸。本文报道了拟杆菌属Pif 1(BaPif 1)与对称双叉双链DNA复合物的结构。两个相互作用的BaPif 1分子结合在部分解绕的dsDNA的每个叉上,并分别与5′臂和3′ ss/dsDNA相互作用。两个BaPif 1分子都是活性解旋酶,它们之间的相互作用可能调节它们的解旋酶活性。BaPif 1与5′臂的结合导致5′ ss/dsDNA连接处的急剧弯曲,从而破坏第一个碱基对。BaPif 1结合到3′ ss/dsDNA连接处,通过稳定未配对的第一个碱基对并接合准备断裂的第二个碱基对来影响双链体解旋。我们的研究结果提供了一个前所未有的洞察两个BaPif 1如何相互协调,以解开分叉的dsDNA。
Pif1 plays multiple roles in maintaining genome stability and preferentially unwinds forked dsDNA, but the mechanism by which Pif1 unwinds forked dsDNA remains elusive. Here we report the structure ofBacteroides spPif1 (BaPif1) in complex with a symmetrical double forked dsDNA. Two interacting BaPif1 molecules are bound to each fork of the partially unwound dsDNA, and interact with the 5′ arm and 3′ ss/dsDNA respectively. Each of the two BaPif1 molecules is an active helicase and their interaction may regulate their helicase activities. The binding of BaPif1 to the 5′ arm causes a sharp bend in the 5′ ss/dsDNA junction, consequently breaking the first base-pair. BaPif1 bound to the 3′ ss/dsDNA junction impacts duplex unwinding by stabilizing the unpaired first base-pair and engaging the second base-pair poised for breaking. Our results provide an unprecedented insight into how two BaPif1 coordinate with each other to unwind the forked dsDNA.
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